Recent investigations suggest that the effects of neural agrin might not be limited to neuromuscular junction formation and maintenance and that other aspects of muscle development might be promoted by agrin. Here we tested the hypothesis that agrin induces a change in the excitability properties in primary cultures of non-innervated human myotubes. Electrical membrane properties of human myotubes were recorded using the whole-cell patch-clamp technique. Cell incubation with recombinant chick neural agrin (1 nM) led to a more negative membrane resting potential. Addition of strophanthidin, a blocker of the Na+/K+ ATPase, depolarized agrin-treated myotubes stronger than control, indicating, in the presence of agrin, a higher contribution of the Na+/K+ ATPase in establishing the resting membrane potential. Indeed, larger amounts of both the a1 and the 2 isoforms of the Na+/K+ ATPase protein were expressed in agrin treated cells. A slight but significant down-regulation of functional apamin-sensitive K+ channels was observed after agrin treatment. These results indicate that neural agrin might act as a trophic factor promoting the maturation of membrane electrical properties during differentiation, confirming the role of agrin as a general promoter of muscle development.
Neural agrin changes the electrical properties of developing human skeletal muscle cells
LORENZON, Paola;SCIANCALEPORE, MARINA
2009-01-01
Abstract
Recent investigations suggest that the effects of neural agrin might not be limited to neuromuscular junction formation and maintenance and that other aspects of muscle development might be promoted by agrin. Here we tested the hypothesis that agrin induces a change in the excitability properties in primary cultures of non-innervated human myotubes. Electrical membrane properties of human myotubes were recorded using the whole-cell patch-clamp technique. Cell incubation with recombinant chick neural agrin (1 nM) led to a more negative membrane resting potential. Addition of strophanthidin, a blocker of the Na+/K+ ATPase, depolarized agrin-treated myotubes stronger than control, indicating, in the presence of agrin, a higher contribution of the Na+/K+ ATPase in establishing the resting membrane potential. Indeed, larger amounts of both the a1 and the 2 isoforms of the Na+/K+ ATPase protein were expressed in agrin treated cells. A slight but significant down-regulation of functional apamin-sensitive K+ channels was observed after agrin treatment. These results indicate that neural agrin might act as a trophic factor promoting the maturation of membrane electrical properties during differentiation, confirming the role of agrin as a general promoter of muscle development.Pubblicazioni consigliate
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